ABSTRACT Melanoma is a highly prevalent cancer that kills patients by metastasizing and destroying distal organs. To date, there are no effective molecular prognostic services available for stratifying patients that are at high risk for metastatic relapse in the adjuvant setting. By stratifying patients into high-risk versus low-risk, therapies could be tailored to those patients that stand to benefit from treatment while sparing those patients who will not relapse and not require costly and potentially harmful therapies. Recent research identified three miRNAs (miR-1908, miR-199a5p, and miR-199a3p) that are overexpressed in metastatic melanoma cells and inhibit expression of apolipoprotein A (ApoE), a metastasis suppressor in melanoma. Cancer-cell secreted ApoE prevents cellular invasion and metastatic endothelial recruitment through engaging melanoma cell LRP1 and endothelial cell LRP8 receptors, respectively. Importantly, elevated expression levels of miRNAs in primary tumor samples performs robustly in stratifying melanoma patients into those with high likelihood from those with very low likelihood for metastatic relapse. Here, Rgenix proposes to develop a prognostic service that predicts the likelihood of melanoma metastasis relapse, based on gene expression analysis of the miRNA/ApoE gene signature in primary tumor slides. The long-term goal is to provide a service that allows all patients diagnosed worldwide with melanoma to be screened and profiled for their risk of developing metastasis. This information will guide clinicians in the critical decision-making for accurate treatment of each patient. To that aim, a protocol for combined expression analysis of the three miRNA by quantitative real-time PCR and ApoE by immunohistochemistry will be developed to stratify patients into those at high versus low risk for metastasis relapse with a >80% positive predictive value. Furthermore, Rgenix will determine the sensitivity and specificity of the assay and establish an optimal threshold for the intensity of each miRNA and ApoE expression that will allow for stratification of high-risk versus low-risk relapsers. A >80% sensitivity and specificity will proof feasibility of the approach and allow proceeding to Phase II. In Phase II the threshold and biomarker combination determined in Phase I will be validated in a large retrospective patient cohort, and improved in order to reach >85% specificity and sensitivity. This threshold will be used to initiate prospective trials in collaboration with Memorial Sloan Kettering Cancer Center. In addition, a CLIA-CAP certified laboratory will be set up and Rgenix will promote the product, and obtain 3rd party reimbursement. Rgenix' prognostic service would prove invaluable for society, which is experiencing increasing incidence of melanoma, and for the clinical community, which is constrained by economic forces to restrict metastasis prevention therapies to patients at highest risk for metastatic relapse.